SN-T 3036-2011出口乳及乳制品中多种有几

中华人民共和国出入境检验检疫行业标准出口乳及乳制品中多种有机氯农药残留量的测定气相色谱-质谱/质谱法2011-09-09发布国家质量监督检验检疫总局I本标准由国家认证认可监督管理委员会提出并归口。本标准起草单位：中华人民共和国重庆出人境检验检疫局、中华人民共和国温州出入境检验检疫1醚、灭蚁灵30种有机氯农药残留量的测定和确证。下列文件对于本文件的应用是必不可少的。凡是注日期的引用文件，仅注日期的版本适用于本文GB/T6682分析实验室用水规格和试验方法4.6无水硫酸钠：650℃灼烧4h,在干燥器内冷却至室温，贮于密封瓶中备用4.8提取液：取适量正己烷(4.1)和丙酮(4.2)按体积比1:1进行混合。4.9凝胶渗透色谱洗脱液：取适量环己烷(4.4)和乙酸乙酯(4.5)按体积比1:1进行混合4.10固相萃取洗脱液：取适量正己烷(4.1)和二氯甲烷(4.3)按体积比5:95进行混合。此溶液在0℃~4℃避光保存。24.13标准中间溶液：取适量的各种标准储备溶液，配制成2μg/mL的标准中间溶液，此溶液在0℃~4.15弗罗里硅土固相萃取小柱：1g/6mL,或相当者，使用前用5mL正己烷(4.1)活化5.3电子天平：感量为0.01g种0.1mg。6.3制样操作过程中应防止样品受到污染或发生残留物含量的变化。准确称取10g试样(精确到0.01g)于50mL/具塞离心管中(奶粉、奶糖加10mL水溶解),加入5g氯化钠，再加入10mL提取液(4、8),用涡旋混匀器振粉1min,4000r/min离心3min,将有机相转7.2净化7.2.1凝胶渗透色谱净化7.2.1.1.2流动相：环己烷-乙酸乙酯(1+1.体积比)。3min。min。7.2.2固相萃取净化7.3.2质谱确证和检测按照7.3.1气相色谱质谱/质谱条件测定样液和标准工作溶液，外标法测定样液中的有机氯农药残留量。样品中待测物残留量应在标准曲线范围之内，如果残留量超出标准曲线范围，应进行适当稀释。在上述色谱条件下，各种有机氯质量色谱峰保留时间参见表A.1。标准品的总离子流色谱图和多在相同实验条件下，样品与标准工作液中待测物质的质量色谱峰相对保留时间在士2.5%以内，并过表1规定的范围，则可判断样品中存在对应的被测物。相对离子丰度允许的相对偏差士50%7.3.3空白试验4SN/T3036—20118结果计算和表述 (1)X,——试样中i组分农药的残留量，单位为微克每千克(pg/kg);9.1测定低限9.2回收率9.2.1液态奶中30种有机氯添加浓度及其回收率数据添加浓度在0.8μg/kg时，回收率在62.2%～105.8%之间；添加浓度在2.0pg/kg时，回收率在65.8%～106.3%之间；添加浓度在10.0pg/kg时，回收率在70.1%～108.8%之间；添加浓度在200.0pg/kg时，回收率在80.0%～106.1%之间。9.2.2奶粉中30种有机氯添加浓度及其回收率数据添加浓度在0.8pg/kg时，回收率在69.6%～100.0%之间；添加浓度在2.0pg/kg时，回收率在68.7%～116.1%之间；添加浓度在10.0μg/kg时，回收率在71.0%～108.7%之间；添加浓度在200.0μg/kg时，回收率在80.1%～110.4%之间。9.2.3酸奶中30种有机氯添加浓度及其回收率数据添加浓度在0.8μg/kg时，回收率在69.7%～105.6%之间；添加浓度在2.0pg/kg时，回收率在70.2%～105.8%之间；添加浓度在10.0pg/kg时，回收率在70.8%～107.5%之间；添加浓度在200.0pg/kg时，回收率在80.4%～109.6%之间。9.2.4冰淇淋中30种有机氯添加浓度及其回收率数据添加浓度在0.8μg/kg时，回收率在69.5%～108.5%之间；添加浓度在2.0μg/kg时，回收率在70.4%～116.1%之间；添加浓度在10.0pg/kg时，回收率在71.5%～108.4%之间；添加浓度在200.0pg/kg时，回收率在80.8%～108.9%之间。59.2.5奶糖中30种有机氯添加浓度及其回收率数据添加浓度在0.8pg/kg时，回收率在71.2%～106.9%之间；添加浓度在2.0μg/kg时，回收率在70.7%～115.7%之间；添加浓度在10.0pg/kg时，回收率在70.5%～109.2%之间；添加浓度在200.0pg/kg时，回收率在80.9%～108.0%之间。6参考质谱条件)GC-MS/MS质谱条件：g)碰撞气：氩气，纯度不低于99.999%;时间窗口化合物保留时间CAS号母离子子离子碰撞能量定性/定量离子丰度比%四氯硝基苯六氯苯608-73-1a-六六六五氯硝基苯林丹β-六六六七氯五氯苯胺1)非商业性声明：附录A所列参考质谱条件是在ThermoTSQQuantumGC型气相色诺申联质谱仪上完成的，此处列出试验用仪器型号仅为提供参考，并不涉及商业目的，鼓励标准使用者尝试不同厂家或型号的仪器。7时间窗口min化合物保留时间minCAS号母离子子离子碰撞能量定性/定量离子丰度比%δ-六六六28.8319-86-8艾氏剂29.9309-00-2甲基五氯苯基硫醚263*33.0～39.5氧化氯丹B23环氧七氯28044-83-9253*反式-氯丹5103-74-2顺式-氯丹5103-71-9266*0,p’-滴滴伊72-55-9α-硫丹88.4959-98-8206°39.5～45.3p,p’-滴滴伊40.5狄氏剂40.560-57-1o,p,-滴滴滴41.1'53-19-0异狄氏剂41.972-20-8o,p’-滴滴涕42.6789-02-6165”p,p’-滴滴滴43.072-54-8β-硫丹43.033213-65-9206*p,p’-滴滴涕43.9789-02-68表A.1(续)时间窗口化合物保留时间CAS号母离子子离子碰撞能量定性/定量离子丰度比%异狄氏剂醛硫丹硫酸盐甲氧滴滴涕异狄氏剂酮灭蚁灵注：“*”定量离子。9(资料性附录)图B.130种有机氮农药标准溶液的总离子流色谱图(4μg/L)RT:16.33-22.08RT:18.47-30.44RT:16.33-22.08RT:18.47-30.44图B.230种有机氯农药标准溶液的选择反应监测(SRM)离子色谱图环氧七氯氢丹(反式/顺式)DDE(o图B.2(续)RelativeRelativeRelativeRelativeRT:43.000-RT:41.96RT:42.6340狄氏剂41424340狄氏剂0-4243AbundanceAbundance0-4441454145AbundanceRT:46.AbundanceRT:46.RelativeRelativeRT:40-甲氧滴滴涕460-异狄氏剂酮RT:44.68RT:46.240-ThisstandardwasdraftedaccordingtoGB/T1.1—2009《Directivesforstandardization—Part1:Structureanddraftingofstandards》,GB/T20001.4—2001《Rulesfordraftingstandards—Part4:Methodsofchemicalanalysis》andSN/T0001—1995《Generalrulesfordraftingthestandardmethodsforthedeterminationofpesticide,veterinarydrugresiduesandbiotoxinsincommoditiesPleasenotethatthisstandardofsomematerialmaybeinvolvedinpatents.ThisstandardissuanceagencydoesnotassumetherThisstandardwasproposedbyandwasunderthechargeoftheCertificationandAccreditationAdministrationofthePeople'sRepublicofChina.ThestandardwasdraftedbyChongqingEntry-ExitInspectionandQuarantiRepublicofChina,WenzhouEntry-ExitInspectionandQuarantineBureauofthChina,ChongqingEngineeringingResearchCenterforImportandExportFoodSafety,ChineseAcade-myofInspectionandQuarantine.ThisstandardwasmainlydraftedbyZhengGuocan,LiuYi,HanChao,ZhangLei,PengTao,LiXianlia-ng,WangGuomin,LiYingguo,CaoShurui,XiCunxian.ThisstandardspecifiesthemethodsofsamplepreparationandqualifiedandquantifieddeterminationbyGC-MS/MSoforganochlorinThisstandardisapplicabletothedeterminationofα-BHC,β-BHC,Lindane,δ-BHC,o,p’-DDT,p,p⁷-DDT,o,p'-DDE,p,p'-DDE,o,p'-DDD,p,p'-DDD,Methoxychlor,Heptachlor,Heptachlorepoxide,Aldrin,Dieldrin,Endrin,Endrin-aldehyde,Endrin-ketone,a-Chlordane(cis),v-Chlordane(trans),Oxychlordane,α-Endosulfan,β-Endosulfan,Endosulfansulfate,Hexachlorobenzene,Tecnazene,Quintozene,Pentachloroaniline,Pentachlorothioanisole,Mirex30organochlorinepesticideresiduesinliquidmilk,milkpowder,yoghurt,icecream,creamcandyet.almilkanddiaryproducts.Thefollowingdocumentsfortheapplicationofthisdocumentisindispensable.Alltilereferencedocumentsdated,whichisonlyapplicabletothedateofnoteversionofthisdocument.Anyonewhodoesn'tdatedreferencedocuments,thelatestversion(includesallofthechanges)applicabletothisstandard.GB/T6682Waterforanalyticallaboratoryuse—SpecificationandtestmethodsTheresiduesoforganochlorinepesticideinthetestsampleareextractedwiththemixedsolutionofhexaneandacetone(1+1,V/V).AfterbeingconcentratedandpurifiedbyGPCandflorisilSPEcolumn,theresiduesaredeterminedbyGC-MS/MS,quantifiedbyexternalstandardmethod.Unlessspecified,allreagentsshouldbeofanalyticalgrade;“Water”istheprimarywateroftheGB/T6682provisioncd.4.1Hexane:HPLCgrade.4.3Dichloromethane:HPLCgrade4.7Sodiumchloride.4.8Extractingsolution:Dilutehexane(4.1)withacetone(4.2)by1:1.4.9GPCelutionsolution:Dilutecyclohexane(4.4)withethylacetate(4.5)by1:1.4.10FlorisilSPEelutionsolution:Dilutehexane(4.1)withdichloromethane(4.3)by5:954.11Organochlorinepesticide:Purity≥95.0%4.12Stockstandardsolution:AccuratelyWeightanadequatoamountofoveryorganochlorinepesticidechloridestandardinanvolumetricflaskanddissolutionof100μg/mL.Thestockstandardsolutioncanbepreservedatthetemperature0℃4.13Intermediatestandardsolution:Accuratelytakenanadequateamountofstockstandardsolu-tion,dilutemixcdstandardsolutionwithhexaneto2ug/mL,Itcanbepreservedatthetemperature0℃~4℃,4.14Standardworkingsolution:Accordingtotherequirement,diluteintermediatestandardsolutionwithhexane,thestandardworkingsolutionwaspreparedimmediatelyasneed.4.15FlorisilSPE:1g/6mLorequivalents.4.16Membranefilter:0.45um,organic5Apparatusandequipment5.1GC-MS/MS:equippedwithElsource,5.2Gelpermeationinchromatography(GPC)5.3Electronicbalance:with0.01gand0.1mgprec5.6Rotaryevaporator.5.7Nitrogenevaporator.5.8Centrifugaltubewithcap:50mL,polytetrafluofoethylene6Samplepreparationandstorage6.1Liquidmilk,yoghurtandicecreamproducts:About100grepresentativesamtakenfromallsamples,putinsuitablecleancontainer.Afterbeing_sealedandlabeled,thesamples6.2Miikpowder,creamcandy/About100greprese6.3CertainmeasuresshouldbetakentoprgventcontamingtlonofthesamplesordecompositionofAccuratelyweigh10gofthetestsample(accurateto0.01g)intoa50mLpolypropylenecentrifugetubewithcap(milkpowder,creamcandydissolvedwith10mLwater),add5gsodiumchlorideand10mLextractant(4.8),oscillatedfor1minwithvortexmixer,thencentrifugeat4000r/minfor3min.Transferthesupernatanttoa100mLrotaryevaporatingflask,theresiduewasextractedwith2×10mLextractant(4.8).Aftercentrifuging,combinethesupernatanttotherotaryevaporatingflask.Evaporatetheextracttodrynessonrotaryevaporatorbelow40℃.Theresidueiswith10.0mLofGPCelutionsolution(4.9)andafterfilteredthroughamilliporemenbranefilterwithanaverageporediameterof0.45μmfilm,thesolutionisreadyforpurifying.SN/T3036—20117.2.1.1.1Purifyingcolumn:400mm×25mm(i.d.),fillingwithpackingofBio-Beads,S-X3,7.2.1.1.2Mobilephase:hexaneandethylacetate(1+1,V/V)7.2.1.1.5Startcollectingtime:10min.7.2.1.1.6Stopcollectingtime:22mTransferthesolutionwaitingforpurified(7.1)to10mLtube,thenpurifiedbyGPC,collecting40℃.ThesolutionisreadyforflorisilSPEcleanupTheflorisilSPEisactivatedwith5mLhexane(4.1).Addedsampletothecolumn,ande8mLflorisilSPEelutionsolution(4.10),thetargetcompoundweretakenoutanddriedtodrynessonrotaryevaporatorbelow40℃,tthroughamilliporemenbranefilterwithanaverageporediameterof0.45μmfilm,thesolutireadyforGC-MS/MSdetermination.7.3.1.1Column:TR-35ms,30m×0.25mm(i.d.)×0.25μm(filmthickness)orequivalents.7.3.1.3Injectiontempe7.3.1.5Interfacetemperatu7.3.1.7Determinationmodel:MS/MS.7.3.1.8MS/MSreferenceconditionslistedtableA.1.7.3.1.10Flowrateofgas:1.2mL/min.7.3.1.11Injectionmode:Splitless.7.3.1.12Injectionvolume:1μL.Accordingtothemethodof7.3.1,detecttheresiduesoforganochlorinepesticideinthetestsamplesolution,thestandardworkingsolution.Theresponseoforganochlorinepesticideshouldbeinthelinearrangeoftheinstrumentaldetection.Iftheresponseisoutofthelinearrange,thesamplebedilutedwithhexane(4.1)tosuitableconcertration.Undertheabovechromatographconditions,thereferenceretentiontimeoforganochlorinepesticideislistedinannexA.RchromatogramofstandardworkingsolutionislistedinannexB.Underthesameconditionsofexperiment,theretentiontimeoftheunknownsampleisthesameasthestandardworkingsolution;thequalificationionsforeverycompoundmustbefound.Forthesameanalysisbatchandthesamecompound,thevariationrangeoftheionratiobetweenthetwodaughterionsfortheunknownsampleandthestandardworkingsocannotbeoutofRelativeintensity>20%～50%>10%～20%PermittedtolerancesbyGC-MS/MSdataprocessororaccordingtotheformula(1): (1)Where:X,-theresiduecontentofcomponentTofposticidesTnthetestSample,ug/kg;c,-theconcentrationofcomponentiofpesticidesinthetestsamplecalculatedbycalibracurve,μg/L;V—thefinalvolumeofsamplesolution,mL;m—thecorrespondingmassoffestsampleinthefinalsample.solution,g.Theblankvalueshallbesubtractedfromtheaboveresultofcalculation.9LimitofquantificationandrecoveryThelimitofquantificationforeveryoriganochlorinepesticideis0.8μg/kg.9.2.1Accordingtotheexperinentaldate,thefortifyingconcentrationoforganochlorinepesticide0.8μg/kg,therecoveryis62.2%～105.8%;2.0μg/kg,therecoveryis65.8%～106.3%;10.0μg/kg,therecoveryis70.1%～108.8%;200.0μg/kg,therecoveryis80.0%～106.1%.9.2.2Accordingtotheexperimentaldate,thefortifyingconcentrationoforganochlorinepesticide0.8ug/kg,therecoveryis69.6%～100.0%;2.0μg/kg,therecoveryis68.7%～116.110.0μg/kg,therecoveryis71.0%～108.7%;200.0μg/kg,therecoveryis80.1%～110.4%.9.2.3Accordingtotheexperimentaldate,thefortifyingconcentrationoforganochlorinepesticide0.8μg/kg,therecoveryis69.7%～105.6%;2.0μg/kg,therecover200.0μg/kg,therecoveryis80.4%～109.6%.9.2.4Accordingtotheexperimentaldate,thefortifyingconcentrationoforganochlorinepesticideinicecreamanditscorrespondingrecoveriesare:0.8μg/kg,therecoveryis69.5%108.5%;2.0μg/kg,therecoveryis70.4%～116.8%;10.0μg/kg,therecoveryis71.5%～108.4%;200.0μg/kg,therecoveryis80.8%～19.2.5Accordingtotheexperinentaldate,thefortifyingcohcentrationoforganochlorinepesticide0.8μg/kg,therecoveryis72.0ug/kg,therecoveryis70.7%～115.7%;10.0ug/kg,therecovery200.0μg/kg,therecoveryis80.9%～108.0%.(Informative)ReferencedconditionsofGC-MS/MS:a)lon